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Salmonella producing β-lactamases has spread rapidly worldwide and poses a serious threat to human and animal health. In this study, we characterized 220 ceftriaxone (CRO)-resistant isolates identified among 3153 Salmonella from humans, animals, food, and water collected in Shanghai, China. They were assessed for antimicrobial susceptibility, phenotypic identification of extended-spectrum β-lactamases (ESBLs), and β-lactamase genes and integrons. CRO resistance in Salmonella increased from 5.0% in 2011 to 8.4% in 2013. Salmonella Enteritidis (45.5%), Salmonella Typhimurium (20.9%) from humans, and Salmonella Indiana (14.5%) from poultry represented the majority of the CRO-resistant isolates. Many isolates were also resistant to other antimicrobials, including nalidixic acid (84.5%), sulfisoxazole (70.5%), and tetracycline (61.8%). Resistance to ciprofloxacin was also found in 33.6% of the isolates. Most isolates (98.2%) were confirmed as ESBL producers. Resistance genes such as blaCTX-M, blaTEM, and blaOXA were detected in 207 (94.1%), 99 (45%), and 53 (24.1%) isolates, respectively. Three types of integron I and one type of integron II were identified in 13 (5.9%) and 2 (0.9%) isolates, respectively. The integrons encompassed 10 different genes: dfrA1/12/17/25, aadA1/2/5, sat2, orfF, and ybeA. Our study underscores concern for increasing CRO resistance, and highlights the widespread ESBL genes in Salmonella enterica.

Die vorliegende Arbeit befasst sich mit Gründungen durch Akademikerinnen und Akademiker mit Migrationshintergrund. Dabei wurden vor allem der Bezug dieser Gründungen zu der Umwelt – dem Gründerökosystem –, in der sie stattfinden, sowie ihre gegenseitigen Wechselwirkungen untersucht. Der Forschungsgegenstand ist die Schnittstelle aus den Bereichen Gründungen, Migrantentum und Hochqualifikation. Der Fokus auf die sehr spezifische Zielgruppe Gründungen durch Akademikerinnen und Akademiker mit Migrationshintergrund füllt eine wichtige Lücke in der bisherigen Forschung. Methodisch gesehen bedient sich diese Arbeit eines theoretischen Bezugsrahmens. Dieser besteht aus der neoinstitutionalistischen Organisationstheorie (Meyer & Rowan 1977), dem Ressourcenabhängigkeitsansatz (Pfeffer & Salancik 1978) sowie dem sechs-dimensionalen Modell des Gründerökosystems (Isenberg 2011). Gründungen durch Akademikerinnen und Akademiker mit Migrationshintergrund müssen ihre interne Ausgestaltung an die Anforderung der institutionellen Umwelt anpassen, um die notwendige Legitimität zu sichern. Dadurch können bei unterschiedlichen Gründungen isomorphe Organisationsstrukturen entstehen. Darüber hinaus können akademische Gründende mit Migrationshintergrund durch interorganisatorische Aktivitäten den Zugang zu nicht-substituierbaren Ressourcen für die Unternehmensgründung bzw. Geschäftsentwicklung ermöglichen bzw. erleichtern. Daher ist die Kombination beider Theorien und des Erklärungsansatzes ein effektives und passendes Analysetool für die vorliegende Forschungsarbeit und schafft sowohl auf Mikro- als auch auf Makroebene für die Leserinnen und Leser ein vollständiges Gesamtbild. Die vorliegende Arbeit beinhaltet nicht nur Daten aus Sekundärquellen und bereits vorhandenen quantitativen Studien im deskriptiven Teil, sondern auch direkte Informationen durch eigene qualitative Untersuchung im empirischen Teil. Dafür wurden insgesamt 23 semistrukturierte Experteninterviews durchgeführt. Durch die Inhaltsanalyse nach Mayring (2014) wurden mehrere Kategorien herausgefiltert; dazu zählen bspw. umweltbezogene Einflussfaktoren auf Legitimität sowie nicht-substituierbare Ressourcen für Gründungen durch Akademikerinnen und Akademiker. Darüber hinaus wurden durch die Empirie einige Hypothesen für weitere quantitative Forschungen in der Zukunft aufgestellt und konkrete Handlungsempfehlungen für die Praxis gegeben., The present paper deals with start-ups by academics with a migration background (in Germany). Above all, the relation of these start-ups to the environment – the entrepreneurial ecosystem – in which they take place, as well as their mutual interactions were examined. The subject of research is the interface between the areas of start-ups, migrants and high qualification. The focus on the very specific target group start-ups by academics with a migration background fills an important gap in previous research. Regarding to the methodological approach, this paper uses a theoretical frame which consists of the theory of neo-institutionalism (Meyer & Rowan 1977), the resource dependence theory (Pfeffer & Salancik 1978) and the six domains model of an entrepreneurship ecosystem (Isenberg 2011). Start-ups by academics with a migration background must adapt their internal structure to the requirements of the institutional environment in order to ensure the necessary legitimacy. This can result in isomorphic organizational structures in different start-ups. In addition, academic founders with a migration background can use interorganizational activities to enable or facilitate access to non-substitutable resources for setting up a company or business development. Therefore, the combination of both theories and the explanatory approach is an effective and suitable analysis tool for the present research work and creates a complete picture for the reader at both the micro and macro level. The present paper contains not only data from secondary sources and already existing quantitative studies in the descriptive part, but also direct information through own qualitative research in the empirical part. A total of 23 semi-structured expert interviews were carried out for this purpose. The content analysis according to Mayring (2014) filtered out several categories; these include, for example, environmental factors influencing legitimacy and non-substitutable resources for start-ups by academics. In addition, some hypotheses for further quantitative research in the future were set up through empirical research and concrete recommendations for action were also given in this paper.

Disease caused by nontyphoidal serotypes of Salmonella enterica is the leading cause of foodborne illness worldwide. Many serotypes have also developed resistance to antimicrobials commonly used for the treatment of salmonellosis. Here we analyze 179 ciprofloxacin-resistant isolates identified among 3680 Salmonella isolated from humans, food, animals, and water collected in Shanghai, China. They were assessed for antimicrobial susceptibility, plasmid-mediated quinolone resistance (PMQR) determinants, and mutations in quinolone resistance determination regions (QRDRs); genetic relatedness was examined using PFGE. Ciprofloxacin resistance in Salmonella increased from 2.3% in 2006 to 5.9% in 2012. Multidrug resistance was common, and most carried mutations in QRDR (97.2%) and PMQR determinants (71.5%). Mutations frequently included changes in gyrA: Ser83Phe (53.6%) and Asp87Asn (35.8%), and in parC: Thr57Ser (53.1%) and Ser80Arg (44.1%). Mutations in parC and parE without changes in gyrA were identified in S. Derby and most S. Thompson. Among PMQR determinants, aac(6')-Ib-cr (62.0%) and oqxA/oqxB (33.5%/33.0%) were most common, and conferred resistance without target mutations in five S. Typhimurium isolates. PFGE analysis revealed that S. Typhimurium, isolated from pork and aquatic products, and S. Indiana, isolated from chicken, were highly similar to isolates from humans, suggesting these products be the major source of ciprofloxacin-resistant infections. Our findings highlight the important role QRDRs and PMQR play in ciprofloxacin resistance of Salmonella, and reveal the potential sources of the pathogen associated with human infections.

A total of 120 Listeria monocytog enes isolates (107 from foods and 13 from humans) in Shanghai, China, were characterized by serogroup typing and virulence genes detection with PCR, antimicrobial susceptibility testing, and molecular subtyping using pulsed-field Gel Electrophoresis (PFGE). Isolates belonged to three L. monocytogenes serogroups, 1/2c, 3c (n = 47, 39.1%) was the most prevalent, followed by 1/2a, 3a (n = 44, 36.7%) and 1/2b, 3b, 7 (n = 29, 24.2%). Although the isolates were all susceptible to most antibiotics tested, 13 isolates were resistant to ceftriaxone and seven to tetracycline. None of the isolates were resistant to multiple antibiotics. The tet (M) genes were detected among tetracycline-resistant isolates. The isolates all harbored virulence genes hly , prfA , plcA , plcB , inlA , inlB , inlI ; 116 isolates (96.7%) were positive for inlC ; 78 (65.0%) for inlG ; 119 (99.2%) for inlJ ; 8 (6.67%) for llsX . A total of 74 distinct patterns were generated for the 119 isolates tested using Apa I and Asc I with Pulsed Field Gel Electrophoresis (PFGE). Three clinical isolates shared the same PFGE patterns with retail meet (chicken, duck, and beef) isolates. These findings indicated that food of animal origin likely serve as a major vehicle in transmitting human listeriosis in Shanghai. Active surveillance for L. monocytogenes is needed for better understanding the epidemiology of the pathogen, and for effective control and prevention of human listeriosis.

Background Internalins are surface proteins that are utilized by Listeria monocytogenes to facilitate its invasion into human intestinal epithelial cells. The expression of a full-length InlA is one of essential virulence factors for L. monocytogenes to cross the intestinal barrier in order to invade epithelial cells. Results In this study, the gene sequences of inlA in 120 L. monocytogenes isolates from food (n = 107) and humans (n = 13) were analyzed. Premature stop codon (PMSC) mutations in inlA were identified in 51 isolates (50 from food and 1 from human). Six mutation types of PMSCs were identified. Among the 51 isolates with PMSCs in inlA, there were 44 serogroup 1/2c, 3c isolates from food, of which seven belonged to serogroups 1/2a, 3a. A total of 153,382 SNPs in 2247 core genes from 42 genomes were identified and used to construct a phylogenetic tree. Serotype 1/2c isolates with inlA PMSC mutations were grouped together. Cell culture studies on 21 isolates showed that the invasion to Caco-2 cells was significantly reduced among isolates with inlA PMSC mutations compared to those without PMSC mutations (P

BACKGROUND Campylobacter is a major cause of bacterial gastroenteritis worldwide. Young children represent a particular age group affected by Campylobacter infection because of their limited diets and weak immune systems. METHODS In this study, a total of 110 Campylobacter (80 Campylobacter jejuni and 30 Campylobacter coli) isolated from children younger than 5 years of age with diarrhea in Shanghai, China in 2011 were examined for their genetic relationship and antimicrobial susceptibility. The presence of virulence genes and its association with invasion potential in Caco-2 cell were also determined. RESULTS Multilocus sequence typing revealed 62 sequence types (STs) under 14 clonal complexes from C. jejuni and 15 STs under 2 clonal complexes from C. coli. High resistance rates among the 110 isolates were observed to nalidixic acid (88.2%), ciprofloxacin (87.3%) and tetracycline (87.3%), followed by ampicillin (30.9%), gentamicin (28.2%), clindamycin (21.8%), erythromycin (21.8%) and chloramphenicol (8.2%). Compared with that of C. jejuni (32.5%), a larger proportion of C. coli (83.3%) were resistant to multiple antimicrobials, including 16 isolates of ST-828 complex resistant to 6 antimicrobials: ciprofloxacin, clindamycin, erythromycin, gentamicin, nalidixic acid and tetracycline. Furthermore, 57 Campylobacter isolates were selected based on their distinct STs and the presence of virulence genes to determine their abilities to adhere to and invade Caco-2 cells. The level of invasion varied widely among isolates and had relatively weak correlation with the genotype data. CONCLUSION Our findings provided baseline data on Campylobacter among young children. Active surveillance of Campylobacter is needed to better understand the epidemiology and antimicrobial resistance trends of this significant pathogen to help control and protect young children from such infections.

Salmonella enterica serovar Stanley is an important serovar that has been increasingly identified in human salmonellosis. The present study aimed to investigate the antimicrobial resistance and molecular typing of 88 Salmonella Stanley strains isolated from humans (diarrhea patients, n = 64; and healthy carrier, n = 1), foods (aquatic products, n = 16; vegetable, n = 1; and pork, n = 1), and environment (waste water, n = 2; and river water, n = 3) in Shanghai, China from 2006 to 2012. Nearly half of the strains were resistant to sulfafurazole (43/88, 48.9%), and many were resistant to streptomycin (35/88, 39.8%), tetracycline (22/88, 25%), and nalidixic acid (19/88, 21.6%). Approximately a quarter of the strains (24/88, 27.3%) were resistant to more than three antimicrobials, and five had ACSSuT resistance type. Six clusters (A-F) were identified by pulsed-field gel electrophoresis (PFGE) with 80% similarity. Interestingly, strains in the same cluster identified by PFGE possessed similar antibiotic resistance patterns. PFGE typing also indicated that aquatic products might serve as a transmission reservoir for Salmonella Stanley infections in humans.

We sequenced 25 isolates of phenotypically multidrug-resistant Salmonella Indiana (n = 11), Typhimurium (n = 8), and Enteritidis (n = 6) using both MinION long-read [SQK-LSK109 and flow cell (R9.4.1)] and MiSeq short-read (Nextera XT and MiSeq Reagent Kit v2) sequencing technologies to determine the advantages of each approach in terms of the characteristics of genome structure, antimicrobial resistance (AMR), virulence potential, whole-genome phylogeny, and pan-genome. The MinION reads were base-called in real-time using MinKnow 3.4.8 integrated with Guppy 3.0.7. The long-read-only assembly, Illumina-only assembly, and hybrid assembly pipelines of Unicycler 0.4.8 were used to generate the MinION, MiSeq, and hybrid assemblies, respectively. The MinION assemblies were highly contiguous compared to the MiSeq assemblies but lacked accuracy, a deficiency that was mitigated by adding the MiSeq short reads through the Unicycler hybrid assembly which corrected erroneous single nucleotide polymorphisms (SNPs). The MinION assemblies provided similar predictions of AMR and virulence potential compared to the MiSeq and hybrid assemblies, although they produced more total false negatives of AMR genotypes, primarily due to failure in identifying tetracycline resistance genes in 11 of the 19 MinION assemblies of tetracycline-resistant isolates. The MinION assemblies displayed a large genetic distance from their corresponding MiSeq and hybrid assemblies on the whole-genome phylogenetic tree, indicating that the lower read accuracy of MinION sequencing caused incorrect clustering. The pan-genome of the MinION assemblies contained significantly more accessory genes and less core genes compared to the MiSeq and hybrid assemblies, suggesting that although these assemblies were more contiguous, their sequencing errors reduced accurate genome annotations. Our research demonstrates that MinION sequencing by itself provides an efficient assessment of the genome structure, antimicrobial resistance, and virulence potential of Salmonella; however, it is not sufficient for whole-genome phylogenetic and pan-genome analyses. MinION in combination with MiSeq facilitated the most accurate genomic analyses.

Introduction: Antimicrobial resistance of Salmonella poses a key threat to public health worldwide. Salmonella Enteritidis (S. Enteritidis) (119 isolates) from children under 10 years old with diarrhea in Shanghai from 2010-2012 were characterized for antimicrobial susceptibility, virulence gene profiles and pulsed field gel electrophoresis (PFGE) patterns. Methodology: The minimum inhibitory concentration for the 119 S. Enteritidis isolates was determined using an agar dilution method. The presence of virulence genes was detected by polymerase chain reaction (PCR). All the isolates with antimicrobial resistance were subjected to PFGE analysis. Results: Among these isolates, 71.4% (85) were resistant to sulfafurazole, 59.7% (71) were resistant to ampicillin, 47.1% (56) were resistant to streptomycin, 7.6% (9) were resistant to ceftiofur and 3.4% (4) were resistant to ceftriaxone. Multidrug resistance (MDR) was found in 63.9% (76) of the isolates; 23 resistance profiles were identified. All isolates harbored the ssaQ and sopE virulence genes in the 16 virulence profiles (VPs); VP1 accounted for 70.59% of the 119 isolates. There were 57 PFGE patterns among the 92 isolates tested, mainly grouped into five clusters (A to E). All of the 76 MDR isolates carried multiple virulence genes. Conclusions: Our study provides useful microbiological data for the successful treatment of S. Enteritidis infections in Shanghai. Although broad spectrum antimicrobials may be useful in the treatment of invasive S. Enteritidis infections, clinicians need to be aware of common microbiological traits, because of the high prevalence of MDR.

Listeria monocytogenes is a foodborne pathogen of global concern because of the high mortality rate among patients. The draft genome sequences of three L. monocytogenes strains isolated from foods are reported here. The availability of these genomes should provide useful information on the genomic diversity of L. monocytogenes isolated from foods in China.

Listeria monocytogenes is a foodborne pathogen of global concern because of the high mortality rate among patients. The draft genome sequences of three L. monocytogenes strains isolated from foods are reported here. The availability of these genomes should provide useful information on the genomic diversity of L. monocytogenes isolated from foods in China.

Salmonella Senftenberg is an important nontyphoidal Salmonella serovar that causes gastrointestinal disease worldwide. In total, 130 Salmonella Senftenberg strains obtained from humans, food, and the environment in Shanghai, People's Republic of China, were characterized for antimicrobial susceptibility and subjected to molecular typing. Our findings indicated that most (96 of 130, 73.8%) of the strains were susceptible to all 13 antimicrobial compounds tested, whereas only two strains (1.5%) were resistant to two antimicrobial compounds. In total, 56 pulsed-field gel electrophoresis profiles were identified, including four main pulsed-field gel electrophoresis profiles (X2, X3, X4, and X5) that showed 95.7% genetic similarity. Our study revealed that the strains of Salmonella Senftenberg from food and the environment shared a high correlation of genetic similarity with those from humans, highlighting the potential links that exist among the strains recovered from different sources in Shanghai.

Terrapins and turtles are known to transmit Salmonella to humans. However, little was known about the occurrence of this pathogen in soft-shelled terrapin that is a popular delicacy in Chinese and other East Asian cuisines. We isolated and characterized 82 (24.4%) isolates of Salmonella from 336 fecal samples of soft-shelled terrapins (51 of 172; 29.7%) and pet turtles (31 of 164; 18.9%) in Shanghai. Salmonella Thompson was the most common serotype (17.1%) among others. Many isolates (84.1%) were resistant to multiple antimicrobials (≥3). Molecular analysis of Salmonella Thompson and Salmonella Typhimurium using pulsed-field gel electrophoresis unveiled a close genetic relationship between several human and terrapin isolates. Our results highlight the risk associated with the handling and consumption of turtles and their role in the spread of Salmonella in the human salmonellosis.

Salmonella Newport (S. Newport) is a major serotype associated with human salmonellosis. A total of 79 S. Newport recovered from humans and other sources in China were characterized for antimicrobial susceptibility, virulence gene profiles and molecular subtypes using pulsed field gel electrophoresis (PFGE). Approximately 63.3% of the isolates were susceptible to all of 16 antimicrobials tested. Nearly one third of the isolates (31.6%) were resistant to sulfisoxazole, 20.3% to tetracycline and 13.9% to nalidixic acid. Twelve isolates (15.2%) were resistant to three or more antimicrobials. Among 10 virulence genes detected, Salmonella pathogenicity island genes avrA, ssaQ, mgtC, siiD, and sopB and fimbrial gene bcfC were present in most of the isolates (93.7% to 100%). Overall, we observed nine distinct virulence gene profiles, three of which (VP1, VP2 and VP3) were most common (86.1%). A total of 56 PFGE patterns were identified and mainly grouped into seven clusters (A to G) with 80% pattern similarity. Isolates from aquatic product shared a high similarity with those from humans in several clusters, highlighting a potential risk of aquatic product as a source of S. Newport that infect humans. Furthermore, there was a strong association between certain PFGE clusters and virulence gene profiles, suggesting virulence subtyping can be a useful epidemiological tool to discriminate S. Newport isolates.

Salmonella enterica serotype Agona (Salmonella Agona) has been among the top 10 serotypes that cause human diarrheal diseases in China. A total of 95 Salmonella Agona (67 from humans, and 28 from animals, food of animal origins, and environmental sources) recovered in Shanghai, China from 2005 to 2011 were subjected to antimicrobial susceptibility testing and molecular subtyping using pulsed-field gel electrophoresis (PFGE). Approximately 68.4% of the Salmonella Agona isolates were pansusceptible to 15 antimicrobial agents tested, and 4 isolates (4.21%) were resistant to at least 3 antimicrobials. PFGE analysis resulted in 41 unique patterns, of which 4 major PFGE patterns (X3, X4, X5, and X6) were grouped together at 96.1% similarity. Isolates of the four patterns included those from food (pork, beef, and chicken) and humans. Our findings showed that the same clones of Salmonella Agona were recovered from human patients and food, and that food of animal origin was potentially a major vehicle of Salmonella Agona in human salmonellosis in Shanghai.

A total of 123 enterotoxigenic Escherichia coli (ETEC) isolates from diarrheal patients from June to December 2012 in Shanghai, China, were examined to determine their genetic relatedness using multilocus sequence typing and pulsed-field gel electrophoresis (PFGE) and for the presence of virulence genes and antimicrobial susceptibility. Twenty-nine sequence types (STs) and 63 PFGE patterns were identified, and results from the 2 subtyping methods correlated well. The 12 isolates of PFGE cluster B all belonged to ST-2332 and were associated with nosocomial neonatal diarrhea. Isolates of a cluster usually had the same set of virulence factors, whereas isolates of different PFGE clusters carried diverse combinations of virulence determinants. Isolates belonging to ST-2332 and ST-182 (n=9) were resistant to at least 6 antimicrobials. Our findings highlighted the need of active surveillance programs for infectious diseases collecting data at both epidemiological and genetic levels that can detect high-risk lineages of pathogens in order to rapidly identify disease outbreaks.